Although a variety of biochemical mechanisms regulate growing cells with respect to their environment, little is known about the maintenance and breakage of dormant state. Disruption of dormancy in bacterial spores (initiation of germination) occurs in response to specific agents, e.g. L-leucine or D-glucose, but the mechanism is unknown. This problem is being studied first by analysis of Bacillus megaterium temperature-sensitive mutants. The metabolism of radioactively labelled initiating agents (C14 - glucose and C14 - leucine) is being measured at permissive and non-permissive temperatures in an attempt to define the temperature-sensitive lesion. Enzymes normally associated with metabolism of initiating agents are being assayed and the necessity of these enzymes for initiation is being tested with mutants that lack a particular enzyme. The spore "lytic" enzyme, that causes hydrolysis of the cortex during initiation, was measured under a variety of conditions. Although it functions during initiation, the lytic activity is not tightly coupled to initiation. We are trying to determine its exact role. Finally, we are studying the proteins removed by various extraction procedures from wild-type and temperature-sensitive spores and we are characterizing a B. megaterium bacteriophage.